Journal: Journal of Cellular and Molecular Medicine

Article Title: RNF7 promotes glioma growth via the PI3K / AKT signalling axis

doi: 10.1111/jcmm.17656

Figure Lengend Snippet: RNF7 was overexpressed in glioma and negatively associated with prognosis. (A) Heat map shows the top upregulated genes in different groups. (B) WB was used to detect the expression of RNF7 protein in tumour and normal tissues. (C) qRT‐PCR was used to detect RNF7 mRNA levels in tumour tissues and normal tissues. (D, E) IHC staining and scoring were used to detect the protein levels of RNF7 in tumour tissues and normal tissues. (F) Log‐rank test of DFS or OS was conducted. Data were presented as mean ± SD from three independent experiments. ** p < 0.01 and *** p < 0.001

Article Snippet: Primary antibodies were used to detect RNF7 (ab181986; Abcam), AKT (ab179463; Abcam), p‐AKT (ab192623; Abcam), PI3K (ab191606; Abcam) and p‐PI3K (ab182651; Abcam).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemistry

Journal: Journal of Cellular and Molecular Medicine

Article Title: RNF7 promotes glioma growth via the PI3K / AKT signalling axis

doi: 10.1111/jcmm.17656

Figure Lengend Snippet: Association of RNF7 expression with clinicopathological characteristics in human gliomas

Article Snippet: Primary antibodies were used to detect RNF7 (ab181986; Abcam), AKT (ab179463; Abcam), p‐AKT (ab192623; Abcam), PI3K (ab191606; Abcam) and p‐PI3K (ab182651; Abcam).

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: RNF7 promotes glioma growth via the PI3K / AKT signalling axis

doi: 10.1111/jcmm.17656

Figure Lengend Snippet: Single factor and multiple factor analysis table

Article Snippet: Primary antibodies were used to detect RNF7 (ab181986; Abcam), AKT (ab179463; Abcam), p‐AKT (ab192623; Abcam), PI3K (ab191606; Abcam) and p‐PI3K (ab182651; Abcam).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: RNF7 promotes glioma growth via the PI3K / AKT signalling axis

doi: 10.1111/jcmm.17656

Figure Lengend Snippet: Overexpression of RNF7 facilitates cell cycle progression and cell growth and inhibits apoptosis. (A) Cell growth curves measured by CCK‐8 between Vector and oeRNF7. (B) RNF7 overexpression facilitated colony formation and histogram quantification (panels). (C) Invasion assays showing that overexpression of RNF7 facilitates cell invasion. The numbers of invading cells are shown. Bars: 50 μm. (D) Typical pictures of live animal imaging between Vector and oeRNF7. Representative pictures of Ki‐67 staining between Vector and oeRNF7. Data were presented as mean ± SD from three independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: Primary antibodies were used to detect RNF7 (ab181986; Abcam), AKT (ab179463; Abcam), p‐AKT (ab192623; Abcam), PI3K (ab191606; Abcam) and p‐PI3K (ab182651; Abcam).

Techniques: Over Expression, CCK-8 Assay, Plasmid Preparation, Imaging, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: RNF7 promotes glioma growth via the PI3K / AKT signalling axis

doi: 10.1111/jcmm.17656

Figure Lengend Snippet: Knockdown of RNF7 suppresses cell cycle progression and cell growth and promotes apoptosis. (A) Cell growth curves measured by CCK‐8 assay between si‐NC and siRNF7#2. (B) RNF7 overexpression inhibited colony formation and histogram quantification (panels). (C) Invasion assays showing that RNF7 knockdown inhibits cell invasion. The numbers of invading cells are shown. Bars: 50 μm. (D) Typical pictures of live animal imaging between si‐NC and siRNF7#2. Representative pictures of Ki‐67 staining between si‐NC and siRNF7#2. Data were presented as mean ± SD from three independent experiments. * p < 0.05 and ** p < 0.01

Article Snippet: Primary antibodies were used to detect RNF7 (ab181986; Abcam), AKT (ab179463; Abcam), p‐AKT (ab192623; Abcam), PI3K (ab191606; Abcam) and p‐PI3K (ab182651; Abcam).

Techniques: CCK-8 Assay, Over Expression, Imaging, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: RNF7 promotes glioma growth via the PI3K / AKT signalling axis

doi: 10.1111/jcmm.17656

Figure Lengend Snippet: RNF7 activates the PI3K/AKT signalling and thereby drives tumour growth. (A) Expression of RNF7, MCM7, PCNA, AKT, p‐AKT, PI3K and p‐PI3K was detected by WB between si‐NC, siRNF7#1 and siRNF7#2. (B) Expression of RNF7, MCM7, PCNA, AKT, p‐AKT, PI3K and p‐PI3K was detected by WB between Vector and oeRNF7.

Article Snippet: Primary antibodies were used to detect RNF7 (ab181986; Abcam), AKT (ab179463; Abcam), p‐AKT (ab192623; Abcam), PI3K (ab191606; Abcam) and p‐PI3K (ab182651; Abcam).

Techniques: Expressing, Plasmid Preparation

Journal: Journal of Cellular and Molecular Medicine

Article Title: RNF7 promotes glioma growth via the PI3K / AKT signalling axis

doi: 10.1111/jcmm.17656

Figure Lengend Snippet: RNF7 influences tumour growth via activating the PI3K/AKT signalling. (A) Expression of RNF7, MCM7, PCNA, AKT, p‐AKT, PI3K and p‐PI3K was measured by WB in different treatment groups. (B) Cell growth curves measured by CCK‐8 assay in different treatment groups. (C) Cell growth measured by colony formation assay in different treatment groups and histogram quantification (panels). Data were presented as mean ± SD from three independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: Primary antibodies were used to detect RNF7 (ab181986; Abcam), AKT (ab179463; Abcam), p‐AKT (ab192623; Abcam), PI3K (ab191606; Abcam) and p‐PI3K (ab182651; Abcam).

Techniques: Expressing, CCK-8 Assay, Colony Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: RNF7 promotes glioma growth via the PI3K / AKT signalling axis

doi: 10.1111/jcmm.17656

Figure Lengend Snippet: RNF7 influences tumour growth via activating the PI3K/AKT signalling. (A) RNF7 overexpression could promote tumour cell migration, invasion and the effect could be attenuated by treatment with PI3K/AKT signalling pathway inhibitors LY294002. (B) Typical pictures of live animal imaging in different treatment groups and representative pictures of Ki‐67 staining in different treatment groups. Data were presented as mean ± SD from three independent experiments. ** p < 0.01 and *** p < 0.001

Article Snippet: Primary antibodies were used to detect RNF7 (ab181986; Abcam), AKT (ab179463; Abcam), p‐AKT (ab192623; Abcam), PI3K (ab191606; Abcam) and p‐PI3K (ab182651; Abcam).

Techniques: Over Expression, Migration, Imaging, Staining

Journal: Disease Markers

Article Title: Pygo1 Regulates the Behavior of Human Non-Small-Cell Lung Cancer via the Wnt/ β -Catenin Pathway

doi: 10.1155/2022/6993994

Figure Lengend Snippet: Pygo1 expression was upregulated in non-small-cell lung cancer. (a) Kaplan-Meier overall survival (OS) curves according to Pygo1 expression levels in database analyzed by Kaplan-Meier Plotter (nlow = 976, nhigh = 949). (b) Pygo1 protein levels in NSCLC tissues (LC) of different pathological grades and adjacent tissues (AD) were detected by western blotting. (c) Protein expression levels were quantified and analyzed by ImageJ and GraphPad Prism software, respectively. The results shown are representative of independent experiments. (d) Quantitative analysis of IHC images using ImageJ. (e) Representative images from immunohistochemistry (IHC) assays of NSCLC of different pathological grades and adjacent tissues. Grade I, grade II, and grade III refer to WHO-IA-LUAD, WHO-IIA-LUAD, and WHO-IIIB-LUSC, respectively. Scale bar: 100 μ m. ns p ≥ 0.05, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: After a 2 h incubation in 5% nonfat dried milk in TBST at room temperature to block nonspecific binding sites, the membranes were incubated overnight at 4°C with primary antibodies against human Pygo1 (Abcam, ab170607), c-Myc (Abcam, ab32072), survivin (Abcam, ab469), E-cadherin (Proteintech, 20874-1-AP), cleaved-caspase-3 (CST, #9664S), CyclinD1 (CST, #55506), RB (CST, #9313S), p16 (Proteintech, 10883-1-AP), p53 (Proteintech, 10442-1-AP), β -catenin (Proteintech, 17565-1-AP), p27Kip1 (Proteintech, 25614-1-AP), CyclinE1 (CST, #20808S), GAPDH (Proteintech, 10494-1-AP), and β -actin (Proteintech, 6008-1-Ig), respectively.

Techniques: Expressing, Western Blot, Software, Immunohistochemistry

Journal: Disease Markers

Article Title: Pygo1 Regulates the Behavior of Human Non-Small-Cell Lung Cancer via the Wnt/ β -Catenin Pathway

doi: 10.1155/2022/6993994

Figure Lengend Snippet: Overexpression of Pygo1 induced G1/S phase transformation of A549 cells and inhibited apoptosis. (a, b) Pygo1 protein was overexpressed in A549 cell lines and its fold change. EV: blank control; Tag2B: negative control; Pygo1: Pygo1 overexpression cells; GAPDH: loading control ( n = 3). (c) Quantitative analysis of cell cycle stage by flow cytometry. (d) Quantitative analysis of cell proliferation detected by CCK-8. (e) Quantitative analysis of apoptosis by flow cytometry. (f, g) Cleaved-caspase-3 protein level detected by western blotting and its fold change. (h, i) Cell cycle-related proteins detected by western blotting and their fold change by ImageJ and GraphPad Prism ( n = 3). ns p ≥ 0.05, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: After a 2 h incubation in 5% nonfat dried milk in TBST at room temperature to block nonspecific binding sites, the membranes were incubated overnight at 4°C with primary antibodies against human Pygo1 (Abcam, ab170607), c-Myc (Abcam, ab32072), survivin (Abcam, ab469), E-cadherin (Proteintech, 20874-1-AP), cleaved-caspase-3 (CST, #9664S), CyclinD1 (CST, #55506), RB (CST, #9313S), p16 (Proteintech, 10883-1-AP), p53 (Proteintech, 10442-1-AP), β -catenin (Proteintech, 17565-1-AP), p27Kip1 (Proteintech, 25614-1-AP), CyclinE1 (CST, #20808S), GAPDH (Proteintech, 10494-1-AP), and β -actin (Proteintech, 6008-1-Ig), respectively.

Techniques: Over Expression, Transformation Assay, Control, Negative Control, Flow Cytometry, CCK-8 Assay, Western Blot

Journal: Disease Markers

Article Title: Pygo1 Regulates the Behavior of Human Non-Small-Cell Lung Cancer via the Wnt/ β -Catenin Pathway

doi: 10.1155/2022/6993994

Figure Lengend Snippet: Pygo1 overexpression in vitro enhanced A549 cell invasion and migration ability and promoted colony formation. (a, b) Transwell experiments and repeated measurement of absorbance (OD) values. (c, d) Wound healing test result visualization and repeated measurement of scratch area changes at different time points. (e) Visualization of colony formation assay results. (f, g) Western blotting analysis of cell lysates with different treatments using rabbit polyclonal antibody to E-cadherin. Quantitative analysis of protein expression levels using ImageJ and GraphPad Prism ( n = 3). ns p ≥ 0.05, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: After a 2 h incubation in 5% nonfat dried milk in TBST at room temperature to block nonspecific binding sites, the membranes were incubated overnight at 4°C with primary antibodies against human Pygo1 (Abcam, ab170607), c-Myc (Abcam, ab32072), survivin (Abcam, ab469), E-cadherin (Proteintech, 20874-1-AP), cleaved-caspase-3 (CST, #9664S), CyclinD1 (CST, #55506), RB (CST, #9313S), p16 (Proteintech, 10883-1-AP), p53 (Proteintech, 10442-1-AP), β -catenin (Proteintech, 17565-1-AP), p27Kip1 (Proteintech, 25614-1-AP), CyclinE1 (CST, #20808S), GAPDH (Proteintech, 10494-1-AP), and β -actin (Proteintech, 6008-1-Ig), respectively.

Techniques: Over Expression, In Vitro, Migration, Colony Assay, Western Blot, Expressing

Journal: Disease Markers

Article Title: Pygo1 Regulates the Behavior of Human Non-Small-Cell Lung Cancer via the Wnt/ β -Catenin Pathway

doi: 10.1155/2022/6993994

Figure Lengend Snippet: Pygo1 upregulation in NSCLC was related to the Wnt signaling pathway. (a, b) Western blotting analysis of lysates with different treatments using rabbit polyclonal antibody to β -catenin, GAPDH was used as a loading control ( n = 3). (c) Representative images of immunohistochemical (IHC) analysis of β -catenin expression levels in NSCLC from different pathological grades and adjacent tissues and (d) quantitative analysis of IHC images using ImageJ. Grade I, grade II, and grade III refer to WHO-IA-LUAD, WHO-IIA-LUAD, and WHO-IIIB-LUSC, respectively. Scale bar: 100 μ m. ns p ≥ 0.05, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (e) Coimmunoprecipitation analysis of TCF4 and β -catenin expression in A549 cells. (f, g) Western blotting analysis of Wnt/ β -catenin signal proteins and quantitative analysis ( n = 3); ns p ≥ 0.05, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (h) Necropsy photographs of mice and xenograft tumors on day 41. The results shown are representative of independent experiments. (i) Tumor weight was measured on day 41 of the experiment (mean ± SE). (j, k) Western blot analysis of the tissues samples and quantitative analysis ( n = 3); ns p ≥ 0.05, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. Equal amounts of lysates were analyzed via western blot analysis and then probed for different proteins.

Article Snippet: After a 2 h incubation in 5% nonfat dried milk in TBST at room temperature to block nonspecific binding sites, the membranes were incubated overnight at 4°C with primary antibodies against human Pygo1 (Abcam, ab170607), c-Myc (Abcam, ab32072), survivin (Abcam, ab469), E-cadherin (Proteintech, 20874-1-AP), cleaved-caspase-3 (CST, #9664S), CyclinD1 (CST, #55506), RB (CST, #9313S), p16 (Proteintech, 10883-1-AP), p53 (Proteintech, 10442-1-AP), β -catenin (Proteintech, 17565-1-AP), p27Kip1 (Proteintech, 25614-1-AP), CyclinE1 (CST, #20808S), GAPDH (Proteintech, 10494-1-AP), and β -actin (Proteintech, 6008-1-Ig), respectively.

Techniques: Western Blot, Control, Immunohistochemical staining, Expressing

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: The predominant methodical workflow in the current study. PDHB: Pyruvate dehydrogenase E1 subunit β; ROC: Receiver operating characteristic; TMB: Tumor mutation burden; MSI: Microsatellite instability; qRT-RCR: Real-time quantitative PCR.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Mutagenesis, Real-time Polymerase Chain Reaction

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Expression levels of pyruvate dehydrogenase E1 subunit β in pan-cancer. A: Comparison of pyruvate dehydrogenase E1 subunit β (PDHB) expression levels in different cancers based on The Cancer Genome Atlas (TCGA) database; B: Expression of PDHB in cancer vs normal tissues in the TCGA database; C: Differential expression of PDHB in cancer and normal tissues in the TCGA joint Genotype Tissue Expression Dataset database; D: Protein levels of PDHB in tumors and normal tissues from the UALCAN database. a P < 0.05, b P < 0.01, c P < 0.001. PDHB: Pyruvate dehydrogenase E1 subunit β; ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma and Endocervical adenocarcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and Paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing, Comparison, Quantitative Proteomics

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Correlation of pyruvate dehydrogenase E1 subunit β expression with tumor stage in multiple cancers and receiver operating characteristic diagnostic analysis. A: Correlation between pyruvate dehydrogenase E1 subunit β (PDHB) gene expression and tumor staging; B: Receiver operating characteristic curve analysis of PDHB in various cancers (AreaUnderROC > 0.7). ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; ESCA: Esophageal carcinoma; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; PAAD: Pancreatic adenocarcinoma; PRAD: Prostate adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing, Diagnostic Assay, Gene Expression

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Mutation frequency of pyruvate dehydrogenase E1 subunit β in pan-cancer. A: Somatic mutation analysis of pyruvate dehydrogenase E1 subunit β (PDHB) in different cancers; B: CBioPortal shows the mutation type and mutation frequency of PDHB sequences.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Mutagenesis

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Correlation analysis of pyruvate dehydrogenase E1 subunit β gene expression with DNA methylation levels in tumor mutation burden, microsatellite instability and pan-cancer. A: Radar plot demonstrating the relationship between tumor mutation burden and pyruvate dehydrogenase E1 subunit β (PDHB) gene expression in various malignancies. Correlation coefficients are indicated by red curves and ranges are indicated by blue values; B: Radar plot showing the relationship between microsatellite instability and PDHB gene expression and various malignancies. Correlation coefficients are shown by blue curves and ranges are shown by green values; C: PDHB methylation levels in the UALCAN database on different cancers. a P < 0.05, b P < 0.01, c P < 0.001. ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma and Endocervical adenocarcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and Paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Gene Expression, DNA Methylation Assay, Mutagenesis, Methylation

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Correlation of pyruvate dehydrogenase E1 subunit β gene expression with stromal and immune scores in different cancers. A: Correlation analysis of pyruvate dehydrogenase E1 subunit β (PDHB) gene expression with immune scores of bladder urothelial carcinoma (BLCA), breast invasive carcinoma (BRCA), brain lower grade glioma (LGG), lung adenocarcinoma (LUAD), lung squamous cell carcinoma, pancreatic adenocarcinoma (PAAD), thyroid carcinoma (THCA), uterine corpus endometrial carcinoma (UCEC); B: Correlation analysis of PDHB gene expression with stromal scores of BLCA, BRCA, LGG, liver hepatocellular carcinoma, LUAD, mesothelioma, ovarian serous cystadenocarcinoma, PAAD, prostate adenocarcinoma, sarcoma, testicular germ cell tumors, THCA, thymoma, UCEC. a P < 0.05, b P < 0.01, c P < 0.001. BRCA: Breast invasive carcinoma; BLCA: Bladder urothelial carcinoma; LGG: Brain lower grade glioma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; PAAD: Pancreatic adenocarcinoma; THCA: Thyroid carcinoma; UCEC: Uterine corpus endometrial carcinoma; BRCA: Breast invasive carcinoma; LIHC: Liver hepatocellular carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PRAD: Prostate adenocarcinoma; SARC: Sarcoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Gene Expression

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Correlation analysis of pyruvate dehydrogenase E1 subunit β expression and tumor immunity. A: TIMER method to analyze the correlation between pyruvate dehydrogenase E1 subunit β (PDHB) and immune cell infiltration; B: CIBERSORT method to analyze PDHB correlation with immune cell infiltration; C: Co-expression analysis of PDHB with tumor chemokines; D: Co-expression analysis of PDHB with tumor chemokine receptors; E: Co-expression analysis of PDHB with immune checkpoint gene. a P < 0.05, b P < 0.01, c P < 0.001. PDHB: Pyruvate dehydrogenase E1 subunit β. ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma and Endocervical adenocarcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and Paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Pyruvate dehydrogenase E1 subunit β expression levels at the single-cell sequencing level. A and B: CancerSEA database demonstrates the correlation of pyruvate dehydrogenase E1 subunit β (PDHB) expression with multiple biological functions in pan-cancer; C: t-Distributed Stochastic Neighbor Embedding plots showing the distribution of PDHB in ovarian serous cystadenocarcinoma, retinoblastoma, and uveal melanoma at the single-cell level. a P < 0.05, b P < 0.01, c P < 0.001. OV: Ovarian serous cystadenocarcinoma; RB: Retinoblastoma; UM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing, Sequencing

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Enrichment analysis of pyruvate dehydrogenase E1 subunit β-associated genes in pan-cancer. A: Interaction network of pyruvate dehydrogenase E1 subunit β (PDHB)-associated biomarkers derived from the BioGRID database; B: GEPIA2.0 showed that PDHB expression was positively correlated with actin related protein 8 (ACTR8), potassium channel tetramerization domain containing 6 (KCTD6), mutl homolog 1 (MLH1), proteasome 26s subunit, non-ATPase 6 (PSMD6), ribonuclease P/MRP subunit p14 (RPP14), and ubiquitin specific peptidase 19 (USP19) genes; C: Heat map showing PDHB expression positively correlated with 6 genes (ACTR8, KCTD6, MLH1, PSMD6, RPP14, USP19); D: Enrichment analysis of PDHB-related genes. PDHB: Pyruvate dehydrogenase E1 subunit β. ACC: Adrenocortical carcinoma; BLCA: Bladder urothelial carcinoma; BRCA: Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma and Endocervical adenocarcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: Head and Neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and Paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Derivative Assay, Expressing, Ubiquitin Proteomics

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Drug sensitivity analysis. Correlation of IC 50 with pyruvate dehydrogenase E1 subunit β expression for different drugs. A: Chelerythrine; B: Nelarabine; C: Fludarabine; D: Fenretinide; E: Lapachone; F: Vorinostat; G: Dasatinib; H: Dolastatin 10. PDHB: Pyruvate dehydrogenase E1 subunit β.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Results of pyruvate dehydrogenase E1 subunit β expression validation. A: Pyruvate dehydrogenase E1 subunit β (PDHB) expression in the human normal hepatocyte cell line (L-O2) and human hepatoma cell lines (SMMC-7721, HepG2, Huh7, H-97); B: PDHB expression in the human normal gastric mucosal cell line (GES-1) and human gastric cancer cell lines (HGC-27, MGC-803, MKN-45); C: PDHB expression in the human normal colonic epithelial cell line (NCM460) and human colon cancer cell lines (SW620, HCT116); D: PDHB expression in the human normal breast cell line (MCF-10A) and breast cancer cell lines (MDA-MB-231, MCF-7); E: PDHB expression in the human normal prostate cell line (RWPE-2) and prostate cancer cell lines (PC-3, 22Rv1 and DU145); F: Validation of PDHB protein expression in the L-O2 and hepatoma cell lines (HepG2, SMMC-7721, Huh7, H-97); G: Quantitative plots; H: Validation of PDHB protein expression in the GES-1 and gastric cancer cell lines (MKN-45, AGS, HGC-27, MGC-803); I: Quantitative plots. a P < 0.05, b P < 0.01, c P < 0.001. PDHB: Pyruvate dehydrogenase E1 subunit β; L-O2: Human normal hepatocyte cell line; GES-1: Gastric mucosal cells; NS: Not significant.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Expressing, Biomarker Discovery

Journal: World Journal of Gastrointestinal Oncology

Article Title: Analysis of the potential biological value of pyruvate dehydrogenase E1 subunit β in human cancer

doi: 10.4251/wjgo.v16.i1.144

Figure Lengend Snippet: Effects of siRNA-pyruvate dehydrogenase E1 subunit β on proliferation, migration and invasion of Huh7 hepatoma cells. A: Transfection efficiency of siRNA-pyruvate dehydrogenase E1 subunit β (PDHB) in Huh7 cell lines; B: The relative expression of mRNA reflects the efficiency of siRNA-PDHB transfection; C: siRNA-PDHB transfection efficiency by protein expression level; D: The CCK-8 method detected the proliferation capacity of Huh7 cell lines; E: Colony formation assay to determine the proliferation capacity of Huh7 cell lines; F: The histogram shows the number of colonies formed; G: Cell scratch assay to detect the migration capacity of Huh7 cell lines; H: Histogram of quantification of cell scratch assay results; I: Transwell method was used to detect the migration and invasion capacity of Huh7 cell lines; J: The histogram shows the number of migrating cells; K: The histogram shows the number of invading cells. a P < 0.05, b P < 0.01, c P < 0.001. PDHB: Pyruvate dehydrogenase E1 subunit β.

Article Snippet: The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 °C shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots.

Techniques: Migration, Transfection, Expressing, CCK-8 Assay, Colony Assay, Wound Healing Assay